Precision sampling of discrete sites identified during in-vivo functional testing in the mammalian heart

Ventricular arrhythmias after myocardial infarction (MI) originate from discrete areas within the MI border zone (BZ), identified during functional electrophysiology tests. Accurate sampling of arrhythmogenic sites for ex-vivo study remains challenging, yet is critical to identify their tissue, cellular and molecular signature. In this study, we developed, validated, and applied a targeted sampling methodology based on individualized 3D prints of the human-sized pig heart. To this end, 3D anatomical models of the left ventricle were created from magnetic resonance imaging and fused with biplane fluoroscopy. Regions of interest for sampling were annotated on the anatomical models, from which we created a unique 3D printed cast with custom slits identifying the annotated regions for sampling. The methodology was validated by retrieving ablation lesions created at predefined locations on the anatomical model. We applied the methodology to sample arrhythmia-vulnerable regions after MI during adrenergic stimulation. A pipeline of imaging was developed to create a 3D high-resolution map of each sample, highlighting the complex interplay of cellular organization, and altered innervation in the BZ.